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Evaluation of cell wall preparations for proteomics: a new procedure for purifying cell walls from Arabidopsis hypocotyls

机译:蛋白质组学细胞壁制备的评估:从拟南芥下胚轴纯化细胞壁的新程序

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摘要

BackgroundThe ultimate goal of proteomic analysis of a cell compartment should be the exhaustive identification of resident proteins; excluding proteins from other cell compartments. Reaching such a goal closely depends on the reliability of the isolation procedure for the cell compartment of interest. Plant cell walls possess specific difficulties: (i) the lack of a surrounding membrane may result in the loss of cell wall proteins (CWP) during the isolation procedure, (ii) polysaccharide networks of cellulose, hemicelluloses and pectins form potential traps for contaminants such as intracellular proteins. Several reported procedures to isolate cell walls for proteomic analyses led to the isolation of a high proportion (more than 50%) of predicted intracellular proteins. Since isolated cell walls should hold secreted proteins, one can imagine alternative procedures to prepare cell walls containing a lower proportion of contaminant proteins.
机译:背景技术对细胞室进行蛋白质组学分析的最终目标应该是彻底鉴定驻留蛋白。排除其他细胞区隔中的蛋白质。达到这样的目标紧密取决于所关注的细胞隔室的隔离程序的可靠性。植物细胞壁具有特殊的困难:(i)在分离过程中缺少周围的膜可能会导致细胞壁蛋白(CWP)的损失;(ii)纤维素,半纤维素和果胶的多糖网络形成了潜在的污染物捕获器,例如作为细胞内蛋白质。几种报道的分离细胞壁进行蛋白质组学分析的方法导致分离出高比例(超过50%)的预测细胞内蛋白。由于分离的细胞壁应容纳分泌的蛋白质,因此可以想象制备含有较低比例的污染蛋白质的细胞壁的替代方法。

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